WO2016061827A1 - 一种苎麻生物脱胶方法 - Google Patents

一种苎麻生物脱胶方法 Download PDF

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WO2016061827A1
WO2016061827A1 PCT/CN2014/089506 CN2014089506W WO2016061827A1 WO 2016061827 A1 WO2016061827 A1 WO 2016061827A1 CN 2014089506 W CN2014089506 W CN 2014089506W WO 2016061827 A1 WO2016061827 A1 WO 2016061827A1
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degumming
ramie
medium
strain
bacterial
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French (fr)
Inventor
余龙江
樊培
杨英
何峰
敖明章
胡祖德
胡振华
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Bubei Jinghua Textile Group Co ltd
Huazhong University of Science and Technology
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Bubei Jinghua Textile Group Co ltd
Huazhong University of Science and Technology
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    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01C—CHEMICAL OR BIOLOGICAL TREATMENT OF NATURAL FILAMENTARY OR FIBROUS MATERIAL TO OBTAIN FILAMENTS OR FIBRES FOR SPINNING; CARBONISING RAGS TO RECOVER ANIMAL FIBRES
    • D01C1/00—Treatment of vegetable material
    • D01C1/04—Bacteriological retting
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus

Definitions

  • the present invention belongs to the field of biotechnology, and more particularly to a method for performing ramie biological degumming using Bacillus sp. HG-28.
  • Ramie single fiber is long, strong in strength, fast in moisture absorption and moisture absorption, and good in thermal conductivity. It is an excellent textile raw material and medical material, and has a wide range of uses. Ramie fiber can be purely spun, or blended with cotton, wool, silk, etc., and its fabric has the advantages of anti-corrosion, antibacterial, breathable, and stiff. China is a big country producing ramie fiber, its output accounts for more than 95% of the world, and it has great demand in the international market. It is a characteristic industry with competitive advantages in the international market.
  • the main bottleneck problem that restricts the development of ramie industry is how to improve the quality of ramie fiber to meet the high-end market demand at home and abroad under the premise of reducing the production cost of ramie fiber and the pollution problem in the production process.
  • ramie contains gum as the total weight 19% to 30% Left and right, before the textile processing, degumming should be carried out.
  • almost all manufacturers use chemical degumming method in the process of ramie degumming. First, soak the ramie with acid, then simmer the ramie with lye. After scouring, the ramie is tortured by the torture machine.
  • the patent discloses a strain capable of producing bast fiber degumming enzyme and its application in ramie and marijuana degumming.
  • the strain has extensive culture conditions, rapid growth and reproduction, and can produce more degumming under the same culture condition. Enzyme, good degumming effect, and It can be completed in 2 to 6 hours, the degumming rate is 95%, the fiber dispersion is 100%, the fiber is not damaged, and there is no environmental pollution.
  • the two-stage seed culture time of the strain is relatively long (accumulated 16 to 28 cumulatively) Hours), after the end of seed culture, it needs to be fermented for another 12 to 18 hours before it can be used for ramie degumming.
  • the production cycle is too long; and the residual rate of fine dry hemp products is high (2.18%), and the breaking strength is too small ( 4.30) cN/dtex) can only meet the requirements of GB/T 20793-2006 second-class products, and is not suitable for large-scale production of high-quality ramie fine dry hemp.
  • Liu Zhengchu et al. in CN101235357B The patent discloses a process for the rapid extraction of ramie fiber by the industrial fermentation of Erwinia.
  • the strain is hobby mannose, and it is secreted for 7 to 9 hours to achieve key enzymes for secreting non-cellulosic degradation (pectinase, ⁇ - The peak of mannanase and xylanase).
  • the bacteria After the bacteria are activated for 5 to 6 hours, they are inoculated on the herbal fiber materials such as ramie and fermented 5 to 6
  • the hourly fermented forage turns blue, stripping and partially degrading the pectin, hemicellulose and part of the lignin associated with the fiber cells, and then removing the non-cellulosic residue attached to the fiber by washing with a high-pressure water column to achieve extraction.
  • the purpose of pure fiber However, the preparation of the activated bacterial liquid is long, the process is cumbersome, and the use of ultrafiltration membrane equipment is required, which is not conducive to the complete activation production process of the seed in the factory.
  • the patent discloses a Bacillus subtilis strain having ricin degumming activity, and the preparation and application thereof, and the system using the strain as the core is used for ramie degumming, and the guar degumming using the system has a short degumming time and a ramie fiber dispersion rate. achieve 100%, degumming rate reaches 90 Above %, the quality of refined hemp reaches the level of chemical degumming.
  • the above several methods have problems such as high price and long incubation time of the culture system, and all of them are only laboratory tests, and there is still a big gap from the actual application in industrial production.
  • the above problems in the prior art mainly lie in the relatively harsh culture conditions of the activation and amplification of the strain, the long incubation time, the long time of degumming, the difficulty in controlling, the excessive consumption of chemicals and water, and the ramie Product quality is not high.
  • the degumming microorganisms and biological degumming technology that are really used in industrial production are still not mature enough to completely solve the bottleneck problem. .
  • the technical problem to be solved by the present invention is to provide a ramie biological degumming method, Solve the problems of serious pollution, strict process conditions, long production cycle and low product quality in the existing ramie degumming industry.
  • a ramie biological degumming method comprising the steps of:
  • degumming bacterial culture using Bacillus sp. HG-28 as a strain, through the strain activation step, expanding the cultivation step, and diluting the step, obtaining the degumming bacteria F for the ramie biological degumming;
  • the ramie raw hemp into the hemp cage is immersed in the degumming bacteria F, the mass ratio of the ramie ramie to the degumming bacterium F is 1: 11 ⁇ 1 : 12 to 0.5 ⁇ 3 m 3 / h
  • the ventilation is introduced into the air.
  • the fermenter is sealed and heated to raise the pressure to 0.1 to 0.15 MPa for 20 to 40 minutes to complete the degumming process.
  • the step S1 is further included before a strain preservation step, wherein the strain storage step is specifically:
  • Bacillus sp. HG-28 was inoculated into a 250 mL Erlenmeyer flask containing 50-100 mL of medium No. 1 and maintained at a temperature of 33-38 ° C for 90-180 r/min. , cultured on a shaker for 3 to 6 h to obtain bacterial liquid A.
  • the bacterial solution A and the glycerin aqueous solution having a volume fraction of 50% were separately transferred into a 1 to 5 mL centrifuge tube in a ratio of 3:2, and uniformly mixed to obtain a bacterial solution B, and stored at 20 ° C for use;
  • the medium No. 1 comprises: 10 g/L peptone, 5 g/L yeast dipping powder, 10 g/L NaCl. After mixing with water and making up to volume, adjust the initial pH to 7 to 7.5.
  • the strain activating step further comprises:
  • the centrifuge tube containing the bacterial solution B moves to room temperature for 5 to 15 minutes, then press the bacterial solution B to 0.1% to 5%.
  • the inoculation amount is inoculated into a 1 L conical flask containing 200-400 mL of medium No. 1 or medium No. 2 and maintained at a temperature of 33 to 38 ° C for 90 to 180 r/min.
  • the rotation speed was cultured on a shaker for 3 to 6 hours to obtain the bacterial liquid C;
  • the second medium comprises: 10 g/L peptone, 5 g/L yeast dipping powder, 5 g/L NaCl. After mixing with water and constant volume, adjust the initial pH value from 6.8 to 7.7.
  • the expanding the culturing step further comprises:
  • the medium No. 3 comprises: 5 to 20 g/L of soybean meal, 5 to 15 g/L of bran, 0 to 5 g/L of NaCl, and 0 to 0.1 g/L of K 2 HPO 4 , 0 to 0.1 g/L of (NH 4 ) 2 SO 4 . After mixing with water and making up to volume, adjust the initial pH to 6.8 to 7.7.
  • the invention has the following beneficial effects: the Bacillus strain Bacillus sp. HG-28 is used for ramie degumming, has low energy consumption and light pollution; the biological degumming time is short, the process is easy to operate, the degumming efficiency is high, and the time of cell culture is high. Short and low cost; the obtained ramie fine dry hemp has high quality, low residual glue rate and high breaking strength, which is superior to the national standard GB/T 20793-2006 "rice fine dry hemp" first-class fine dry hemp appearance and various technical indicators Large-scale production has broad prospects.
  • the Bacillus sp. HG-28 used in the present invention is isolated from the soil of the castor plantation in Xian'an District of Xianning County by using a selective medium, and is identified as Bacillus by 16 S rDNA sequence analysis and construction of a phylogenetic tree, and It is named Bacillus sp. HG-28.
  • Bacillus sp. HG-28 was submitted to the China Center for Type Culture Collection (CCTCC, address Wuhan University, Wuhan, Hubei province, China) on July 2, 2013. The deposit number is CCTCC No: M 2013308.
  • Strain activation procedure Move the centrifuge tube containing the bacterial solution B to room temperature for 5 minutes, then press the bacterial solution B to 0.1%.
  • the inoculation amount was inoculated into a 1 L Erlenmeyer flask containing 200 No. 1 medium, and maintained at 180 r/min at a temperature of 38 ° C for 6 h on a shaker to obtain a bacterial solution C.
  • the formulation of No. 1 medium is: 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder and 10 g/L NaCl, and the initial pH value is adjusted to 7 .
  • Inoculum C was inoculated into a 50 L seed tank containing 35 L of No. 2 medium at a rate of 5%, maintaining a rotational speed of 90 r/min and 5 L/ at a temperature of 33 °C.
  • the aeration of min was cultured for 5 h to obtain the bacterial solution D.
  • the bacterial solution D was inoculated into a 1 t seed tank containing 700 kg of the second medium, and kept at a temperature of 38 ° C for 60 r / min.
  • the rotation speed and the aeration of 0.5 m 3 /h were cultured for 2 hours to obtain the bacterial solution E.
  • the formulation of the second medium was: 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder, 5 g/L of NaCl and adjust the initial pH to 6.8.
  • the dilution step includes: injecting 5.5t of water into the fermenter, maintaining the temperature at 34 ° C, adjusting the pH value to 6.8, and the bacterial solution E Inject the mixture into the fermenter at a volume fraction of 2%, and mix evenly to obtain the degumming bacteria F.
  • Biological degumming The ramie raw hemp loaded into the cage is immersed in the degumming bacteria F.
  • the mass ratio of the ramie ramie to the degumming bacterium F is 1:1:1, and the air is introduced into the air at a flow rate of 3 m 3 /h.
  • the fermenter was sealed and heated to raise the pressure to 0.15 MPa for 40 min to end the degumming process.
  • the fineness of the ramie single fiber after degumming is 5.26 dtex.
  • the breaking strength of the bundle fiber is 5.02 cN/dtex, the residual rubber ratio is 1.75%, the whiteness is 51 degree, the pH value is 6.89, and the moisture regain rate is 9.13%.
  • Strain activation procedure Move the centrifuge tube containing the bacterial solution B to room temperature for 15 min, then press the bacterial solution B at 5%.
  • the inoculation amount was inoculated into a 1 L conical flask containing 400 mL of No. 1 medium, and maintained at 90 r/min at 33 ° C for 3 h to obtain a bacterial solution C.
  • the formulation of No. 1 medium is: 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder and 10 g/L NaCl, and the initial pH value is adjusted to 7.2. .
  • Inoculum C was inoculated into a 50 L seed tank containing 25 L of No. 2 medium at a temperature of 38 ° C, maintained at 180 r/min and 30 L/ at a temperature of 38 ° C.
  • the ventilation rate of min was cultured for 2 h to obtain the bacterial liquid D;
  • the medium of the second medium was prepared by mixing 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder, and 5 g/L NaCl. And adjust the initial pH to 7.2.
  • inoculum D was inoculated into a 1 t seed tank containing 300 kg of medium No.
  • the medium of the third medium is prepared by mixing 5 g/L of soybean meal, 15 g/L of bran, 5 g/L of NaCl per unit volume of water, and adjusting the initial pH value to 6.8. .
  • the dilution step includes: injecting 5.5t of water into the fermenter, keeping the temperature at 37 ° C, adjusting the pH value to 7.5, and the bacterial solution E Inject the fermenter in a volume fraction of 6% and mix well to obtain the degumming bacteria F.
  • Biological degumming The ramie raw hemp loaded into the cage is immersed in the degumming bacteria F.
  • the mass ratio of the ramie ramie to the degumming bacterium F is 1: 12, and the air is introduced into the air at a ventilation of 0.5 m 3 /h.
  • the fermenter was sealed and heated to raise the pressure to 0.1 MPa for 20 min to complete the degumming process.
  • the fineness of the ramie single fiber after degumming is 5.19 dtex.
  • the breaking strength of the bundle fiber is 5.05 cN/dtex, the residual gel ratio is 1.63%, the whiteness is 49 degrees, the pH is 7.01, and the moisture regain is 9.06%.
  • Strain activation procedure Move the centrifuge tube containing the bacterial solution B to room temperature for 10 min, then press the bacterial solution B at 3%.
  • the inoculation amount was inoculated into a 1 L conical flask containing 300 mL of No. 2 medium, and maintained at a temperature of 37 ° C for 120 h/min, and cultured on a shaker for 4 h to obtain a bacterial solution.
  • No. 2 medium is formulated by mixing 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder, 5 g/L NaCl, and adjusting the initial pH value to 7.7. .
  • Expanding the culture step Inoculating the bacterial solution C into a 50 L seed tank containing 30 L of No. 2 medium at a temperature of 37 ° C, maintaining a rotation speed of 120 r/min and 20 L/min aeration, cultured for 4 h, to obtain bacterial solution D; then inoculum D was inoculated into a 1 t seed tank containing 500 kg of medium No. 3, maintained at a temperature of 37 ° C, maintained at 120 r / The rotation speed of min and the ventilation of 1.5 m 3 /h were cultured for 4 h to obtain the bacterial liquid E.
  • the formulation of the third medium was: 20 g/L of soy meal, 5 bran, 0.1 per unit volume of water. g/L of K 2 HPO 4 , 0.1 g/L of (NH 4 ) 2 SO 4 , and adjusting the initial pH to 7.7.
  • the dilution step includes: injecting water into the fermenter, keeping the temperature at 36 ° C, adjusting the pH value to 7.0, and pressing the bacterial solution E to 4%.
  • the volume fraction is injected into the fermenter and mixed evenly to obtain the degummed bacterial liquid F.
  • Biological degumming The ramie ramie loaded into the cage is immersed in the degumming bacterium F, and the mass ratio of the ramie ramie to the degumming bacterium F is 1: 12, and the air is introduced into the air at 1.5 m 3 /h. After degumming for 6 h, the fermenter was sealed and heated to raise the pressure to 0.12 MPa for 30 min to complete the degumming process.
  • the fineness of the ramie single fiber after degumming is 5.12 dtex.
  • the breaking strength of the bundle fiber is 5.15 cN/dtex, the residual gel ratio is 1.53%, the whiteness is 51 degrees, the pH is 6.98, and the moisture regain is 9.21%.

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Abstract

本发明涉及一种苎麻生物脱胶方法，包括如下步骤：使用芽孢杆菌Bacillus sp. HG-28作为菌种，经过菌种活化步骤、扩大培养步骤、稀释步骤，获得用于苎麻生物脱胶的脱胶菌液F ，然后将苎麻原麻浸没于脱胶菌液F中进行生物脱胶。实施本发明，能耗较小、污染较轻；生物脱胶时间短、过程易操作、脱胶效率高；菌体培养的时间短、成本低；得到的苎麻精干麻品质高、残胶率低、断裂强度高，优于国家标准GB/T 20793-2006一级精干麻的外观和各项技术指标的要求；规模化生产前景广阔。

Description

一种苎麻生物脱胶方法 技术领域
本发明属于生物技术领域，更具体的说，涉及一种使用 芽孢杆菌 Bacillus sp. HG-28 进行 苎麻生物脱胶的方法。
背景技术
苎麻， 学名 Boehmeria nivea (L.) Gaud. ，是一种提供重要纺织纤维的经济作物。 苎麻单纤维长、强度大、吸湿散湿快、导热性好，是优良纺织原料及医用材料，用途广泛。苎麻纤维可以纯纺，也可与棉、毛、丝等混纺，其织物具有防腐、抑菌、透气、挺括等优点。我国是苎麻纤维生产大国，其产量占世界 95% 以上，在国际市场具有重大需求，是我国具有国际市场竞争优势的特色产业。
目前，制约苎麻产业发展存在的主要瓶颈问题是如何在降低苎麻纤维生产成本与生产过程的污染问题的前提下，提高苎麻纤维的品质，以满足国内外高端市场需求。由于苎麻含胶质占总重量 19% ～ 30% 左右，在纺织加工前要进行脱胶，目前，几乎所有厂家在苎麻脱胶过程中都采用化学脱胶方法，首先用酸浸泡苎麻，然后用碱液煮练苎麻，煮练后的苎麻通过拷打机拷打后，再用漂白剂漂洗、酸中和，然后水洗、脱水、给油、烘干得到成品苎麻精干麻。苎麻的化学脱胶需要使用大量酸、碱，产生的废水量大且很难处理，对环境会造成大量污染。因此，目前不少苎麻初加工企业由于生产废水污染水体、不能达标排放被迫关停，以致苎麻原料需求减小，农民种麻收入得不到保障，种麻积极性受到打击，整个产业面临困境。所以，苎麻脱胶过程中的节能减排技术问题已成为制约苎麻产业可持续发展的瓶颈。
近年来，为了减少或根治脱胶造成的污染，已公认利用微生物进行苎麻脱胶是未来主要发展方向，相继开展了苎麻生物脱胶技术研究和相关工艺探索。
刘自鎔等在 CN1371990A 号专利中公开了一株可产生韧皮纤维脱胶酶的菌株及其在苎麻、大麻脱胶中的应用，该菌株具有培养条件粗放、生长繁殖快，在同一培养条件下可产生脱胶所需的多种酶，脱胶效果好，且 2 ～ 6 小时即可完成，脱胶率达到 95% ，纤维分散度 100% ，不损伤纤维，无环境污染。但是该菌株两级种子培养时间比较长（累计达 16 ～ 28 小时），种子培养结束后还需再发酵 12 ～ 18 小时后才能用于苎麻脱胶，生产周期过长；且精干麻产品残胶率偏高（ 2.18% ），断裂强度偏小（ 4.30 cN/dtex ），只能达到 GB/T 20793-2006 二级品的要求，不适合用于高品质苎麻精干麻大规模生产。
刘正初等在 CN101235357B 号专利中公开了一种欧文氏杆菌工厂化发酵快速提取苎麻纤维工艺。该菌株嗜好甘露糖，纯培养 7 ～ 9 小时达到分泌非纤维素降解关键酶（果胶酶、 β- 甘露聚糖酶和木聚糖酶）的高峰。将其菌剂活化 5 ～ 6 小时后接种到苎麻等草本纤维原料上发酵 5 ～ 6 小时的发酵草料变成蓝色，剥离和部分降解伴生于纤维细胞间的果胶、半纤维素及部分木质素，再借助高压水柱冲洗可除去附着在纤维上的非纤维素残留物，达到提取纯净纤维的目的。但是其活化菌液制备时间较长、过程较繁琐、且需要使用超滤膜设备，不利于在工厂里进行种子的全套活化生产程序。
曹军卫等在 CN1177003C 号专利中公开了一株新的嗜碱芽孢杆菌菌株及其在苎麻脱胶中的应用。该发明提供的技术具有脱胶周期短，脱胶效率高，菌种不易被污染，酶活力稳定，处理成本低，纤维质量好等优点。吴瑜等在 CN101270343A 号专利中公开了一种苎麻脱胶菌。该发明苎麻脱胶菌的脱胶能力有质的提高，对麻纤维无损伤，经过脱胶后杂质几乎全部脱掉，脱胶速度至少提高一倍。张兴群等在 CN101654660B 号专利中公开了一种具有苎麻脱胶活性的枯草芽孢杆菌菌株及其制备和应用，利用以该菌株为核心的体系用于苎麻脱胶，利用该体系进行苎麻脱胶具有脱胶时间短，苎麻纤维分散率达到 100 ％，脱胶率达到 90 ％以上，精干麻质量达到化学脱胶水平。上述几种方法存在培养体系价格高、培养时间长等问题，且均只是实验室小试，离实际运用于工业生产尚有较大差距。
以上现有技术存在的问题主要在于所述菌株活化与扩增环节培养条件相对严苛、培养时间较长，脱胶环节存在时间长、难于控制、化学品与水的消耗量偏大，苎麻精干麻产品品质不高。目前，真正用于工业生产的脱胶微生物与生物脱胶技术尚不成熟，未能完全解决存在的瓶颈问题 。
发明内容
本发明所要解决的技术问题在于： 提供 一种苎麻生物脱胶方法 ， 解决现有苎麻脱胶工业存在的污染严重、工艺条件严苛、生产周期长、产品品质不高的问题。
本发明解决上述问题的技术方案在于：
提供一种苎麻生物脱胶方法，所述方法包括如下步骤：
S1 、脱胶菌液培养：使用 芽孢杆菌 Bacillus sp. HG-28 作为菌种，经过 菌种 活化步骤、扩大培养步骤、稀释步骤，获得用于 苎麻生物脱胶的脱胶菌液 F ；
S2 、生物脱胶：将 装入麻笼的苎麻原麻浸没于 脱胶 菌液 F 中， 苎麻 原麻 与脱胶菌液 F 的质量比为 1 ∶ 11 ～ 1 ∶ 12 ，以 0.5 ～ 3 m3/h 的通气量通入空气， 生物脱胶 4 ～ 12 h 后，将发酵罐密封，加热使压力升至 0.1 ～ 0.15 MPa ，保持 20 ～ 40 min ，结束脱胶过程。
在本发明的 苎麻生物脱胶方法中，所述步骤 S1 之前还包括 菌种保存步骤，所述菌种保存步骤具体为：
将芽孢杆菌 Bacillus sp. HG-28 菌种接种到装有 50 ～ 100 mL 一号培养基的 250 mL 锥形瓶中，在 33 ～ 38°C 的温度下、保持 90 ～ 180 r/min 的转速，于摇床上培养 3 ～ 6 h ，得到菌液 A 。将菌液 A 与体积分数为 50% 的甘油水溶液按 3:2 的比例分别移入 1 ～ 5 mL 的离心管内混和均匀，得到菌液 B ， − 20°C 条件下保存备用；
其中，所述一号培养基包括： 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 10 g/L 的 NaCl 。与水混和、定容后，调节初始 pH 值为 7 ～ 7.5 。
在本发明的 苎麻生物脱胶方法中，所述菌种 活化步骤进一步 包括：
将保存有菌液 B 的离心管移至室温下保持 5 ～ 15 min ，然后将菌液 B 按 0.1% ～ 5% 的接种量接种到装有 200 ～ 400 mL 一号培养基或二号培养基的 1 L 锥形瓶中，在 33 ～ 38°C 的温度下、保持 90 ～ 180 r/min 的转速，于摇床上培养 3 ～ 6 h ，得到菌液 C ；
其中， 所述二号培养基包括： 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 5 g/L 的 NaCl 。与水混和、定容后，调节初始 pH 值为 6.8 ～ 7.7 。
在本发明的 苎麻生物脱胶方法中，所述 扩大培养步骤进一步 包括：
将所述菌液 C 按 1% ～ 5% 的接种量接种到装有 25 ～ 35 L 二号培养基的 50 L 种子罐中，在 33 ～ 38°C 的温度下、保持 90 ～ 180 r/min 的转速和 5 ～ 30 L/min 的通气量，培养 2 ～ 5 h ，得到菌液 D ；
然后将菌液 D 接种到装有 300 ～ 700 kg 二号培养基或三号培养基的 1 t 种子罐中，在 33 ～ 38°C 的温度下、保持 60 ～ 150 r/min 的转速和 0.5 ～ 3 m3/h 的通气量，培养 2 ～ 5 h ，得到菌液 E ；
其中，所述三号培养基包括： 5 ～ 20 g/L 的黄豆粕粉、 5 ～ 15g/L 的麸皮、 0 ～ 5 g/L 的 NaCl 、 0 ～ 0.1 g/L 的 K2HPO4 、 0 ～ 0.1 g/L 的 (NH4)2SO4 。与水混和、定容后，调节初始 pH 值为 6.8 ～ 7.7。
实施本发明，具有如下有益效果：芽孢杆菌菌株 Bacillus sp. HG-28 用于苎麻脱胶，能耗较小、污染较轻；生物脱胶时间短、过程易操作、脱胶效率高；菌体培养的时间短、成本低；得到的苎麻精干麻品质高、残胶率低、断裂强度高，优于国家标准 GB/T 20793-2006 《苎麻精干麻》一级精干麻的外观和各项技术指标的要求；规模化生产前景广阔。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
本发明使用的芽孢杆菌 Bacillus sp. HG-28 是使用选择培养基从 咸宁咸安区苎麻种植园土壤中分离得到的，通过 16S rDNA 序列分析和构建系统发育树被鉴定为芽孢杆菌，并将其命名为 Bacillus sp. HG-28 。 Bacillus sp. HG-28 已于 2013 年 7 月 2 号提交给中国典型培养物保藏中心（简称 CCTCC ，地址在中国湖北省武汉市武汉大学）保藏，保藏编号为 CCTCC No: M 2013308 。
实施例 1
菌种 活化步骤 ： 将保存有菌液 B 的离心管移至室温下保持 5min ，然后将菌液 B 按 0.1% 的接种量接种到装有 200 一号培养基的 1 L 锥形瓶中，在 38°C 的温度下、保持 180 r/min 的转速，于摇床上培养 6 h ，得到菌液 C ；一号培养基的配方是：单位体积的水中混和有 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉与 10 g/L 的 NaCl ，并调节初始 pH 值为 7 。
扩大培养步骤 ： 将菌液 C 按 5% 的接种量接种到装有 35 L 二号培养基的 50 L 种子罐中，在 33°C 的温度下、保持 90 r/min 的转速和 5 L/min 的通气量，培养 5 h ，得到菌液 D ；然后将菌液 D 接种到装有 700 kg 二号培养基的 1 t 种子罐中，在 38°C 的温度下、保持 60 r/min 的转速和 0.5 m3/h 的通气量，培养 2h ，得到菌液 E ；二号培养基的配方是：单位体积的水中混和有 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 5 g/L 的 NaCl ，并调节初始 pH 值为 6.8 。
稀释步骤 包括： 将 5.5t 水注入发酵罐中，保温 34°C ，调节 pH 值为 6.8 ，将菌液 E 按 2% 的体积分数注入发酵罐中，混和均匀，得到脱胶菌液 F 。
生物脱胶：将 装入麻笼的苎麻原麻浸没于 脱胶 菌液 F 中， 苎麻 原麻 与脱胶菌液 F 的质量比为 1 ∶ 11 ，以 3 m3/h 的通气量通入空气， 生物脱胶 12 h 后，将发酵罐密封，加热使压力升至 0.15 MPa ，保持 40 min ，结束脱胶过程。
生物脱胶完成后，经过拷麻、漂白、给油、烘干等工序， 经检测，脱胶后的苎麻单纤维细度为 5.26 dtex 、束纤维断裂强度 5.02 cN/dtex 、残胶率 1.75% 、白度 51 度、 pH 值 6.89 、回潮率 9.13% 。
实施例 2
菌种 活化步骤 ： 将保存有菌液 B 的离心管移至室温下保持 15 min ，然后将菌液 B 按 5% 的接种量接种到装有 400 mL 一号培养基的 1 L 锥形瓶中，在 33°C 的温度下、保持 90 r/min 的转速，于摇床上培养 3 h ，得到菌液 C ；一号培养基的配方是：单位体积的水中混和有 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉与 10 g/L 的 NaCl ，并调节初始 pH 值为 7.2 。
扩大培养步骤 ： 将菌液 C 按 1% 的接种量接种到装有 25 L 二号培养基的 50 L 种子罐中，在 38°C 的温度下、保持 180 r/min 的转速和 30 L/min 的通气量，培养 2 h ，得到菌液 D ；二号培养基的配方是：单位体积的水中混和有 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 5 g/L 的 NaCl ，并调节初始 pH 值为 7.2 。然后将菌液 D 接种到装有 300 kg 三号培养基的 1 t 种子罐中，在 33°C 的温度下、保持 150 r/min 的转速和 3 m3/h 的通气量，培养 5 h ，得到菌液 E ；三号培养基的配方是：单位体积的水中混和有 5 g/L 的黄豆粕粉、 15 g/L 麸皮、 5 g/L 的 NaCl ，并调节初始 pH 值为 6.8 。
稀释步骤 包括： 将 5.5t 水注入发酵罐中，保温 37°C ，调节 pH 值为 7.5 ，将菌液 E 按 6% 的体积分数注入发酵罐中，混和均匀，得到脱胶菌液 F 。
生物脱胶：将 装入麻笼的苎麻原麻浸没于 脱胶 菌液 F 中， 苎麻 原麻 与脱胶菌液 F 的质量比为 1 ∶ 12 ，以 0.5 m3/h 的通气量通入空气， 生物脱胶 4 h 后，将发酵罐密封，加热使压力升至 0.1 MPa ，保持 20 min ，结束脱胶过程。
生物脱胶完成后，经过拷麻、漂白、给油、烘干等工序， 经检测，脱胶后的苎麻单纤维细度为 5.19 dtex 、束纤维断裂强度 5.05 cN/dtex 、残胶率 1.63% 、白度 49 度、 pH 值 7.01 、回潮率 9.06% 。
实施例 3
菌种 活化步骤 ： 将保存有菌液 B 的离心管移至室温下保持 10 min ，然后将菌液 B 按 3% 的接种量接种到装有 300 mL 二号培养基的 1 L 锥形瓶中，在 37°C 的温度下、保持 120 r/min 的转速，于摇床上培养 4 h ，得到菌液 C ；二号培养基的配方是：单位体积的水中混和有 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 5 g/L 的 NaCl ，并调节初始 pH 值为 7.7 。
扩大培养步骤 ： 将所述菌液 C 按 3% 的接种量接种到装有 30 L 二号培养基的 50 L 种子罐中，在 37°C 的温度下、保持 120 r/min 的转速和 20 L/min 的通气量，培养 4 h ，得到菌液 D ；然后将菌液 D 接种到装有 500 kg 三号培养基的 1 t 种子罐中，在 37°C 的温度下、保持 120 r/min 的转速和 1.5 m3/h 的通气量，培养 4 h ，得到菌液 E ；三号培养基的配方是：单位体积的水中混和有 20 g/L 的黄豆粕粉、 5 麸皮、 0.1 g/L 的 K2HPO4 、 0.1 g/L 的 (NH4)2SO4 ，并调节初始 pH 值为 7.7 。
稀释步骤 包括： 将水注入发酵罐中，保温 36°C ，调节 pH 值为 7.0 ，将菌液 E 按 4% 的体积分数注入发酵罐中，混和均匀，得到脱胶菌液 F 。
生物脱胶：将 装入麻笼的苎麻原麻浸没于 脱胶 菌液 F 中， 苎麻 原麻 与脱胶菌液 F 的质量比为 1 ∶ 12 ，以 1.5 m3/h 的通气量通入空气， 生物脱胶 6 h 后，将发酵罐密封，加热使压力升至 0.12 MPa ，保持 30 min ，结束脱胶过程。
生物脱胶完成后，经过拷麻、漂白、给油、烘干等工序， 经检测，脱胶后的苎麻单纤维细度为 5.12 dtex 、束纤维断裂强度 5.15 cN/dtex 、残胶率 1.53% 、白度 51 度、 pH 值 6.98 、回潮率 9.21% 。

Claims (5)

  1. 一种苎麻生物脱胶方法，其特征在于，所述方法包括如下步骤：
    S1 、脱胶菌液培养：使用 芽孢杆菌 Bacillus sp. HG-28 作为菌种，经过 菌种 活化步骤、扩大培养步骤、稀释步骤，获得用于 苎麻生物脱胶的脱胶菌液 F ；
    S2 、生物脱胶：将 装入麻笼的苎麻原麻浸没于 脱胶 菌液 F 中， 苎麻 原麻 与脱胶菌液 F 的质量比为 1 ∶ 11 ～ 1 ∶ 12 ，以 0.5 ～ 3 m3/h 的通气量通入空气， 生物脱胶 4 ～ 12 h 后，将发酵罐密封， 加热使压力升至 0.1 ～ 0.15 MPa ，保持 20 ～ 40 min ，结束脱胶过程。
  2. 据权利要求 1 所述的 苎麻生物脱胶方法，其特征在于，所述步骤 S1 之前还包括 菌种保存步骤，所述菌种保存步骤具体为：
    将芽孢杆菌 Bacillus sp. HG-28 菌种接种到装有 50 ～ 100 mL 一号培养基的 250 mL 锥形瓶中，在 33 ～ 38°C 的温度下、保持 90 ～ 180 r/min 的转速，于摇床上培养 3 ～ 6 h ，得到菌液 A ;将菌液 A 与体积分数为 50% 的甘油水溶液按 3:2 的比例分别移入 1 ～ 5 mL 的离心管内混和均匀，得到菌液 B ， − 20°C 条件下保存备用；
    其中，所述一号培养基包括： 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 10 g/L 的 NaCl 。
  3. 根据权利要求 2 所述的 苎麻生物脱胶方法，其特征在于，所述菌种 活化步骤进一步 包括：
    将保存有菌液 B 的离心管移至室温下保持 5 ～ 15 min ，然后将菌液 B 按 0.1% ～ 5% 的接种量接种到装有 200 ～ 400 mL 一号培养基或二号培养基的 1 L 锥形瓶中，在 33 ～ 38°C 的温度下、保持 90 ～ 180 r/min 的转速，于摇床上培养 3 ～ 6 h ，得到菌液 C ；
    其中， 所述二号培养基包括： 10 g/L 的蛋白胨、 5 g/L 的酵母浸粉、 5 g/L 的 NaCl 。
  4. 根据权利要求 3 所述的 苎麻生物脱胶方法，其特征在于，所述 扩大培养步骤进一步 包括：
    将所述菌液 C 按 1% ～ 5% 的接种量接种到装有 25 ～ 35 L 二号培养基的 50 L 种子罐中，在 33 ～ 38°C 的温度下、保持 90 ～ 180 r/min 的转速和 5 ～ 30 L/min 的通气量，培养 2 ～ 5 h ，得到菌液 D ；
    然后将菌液 D 接种到装有 300 ～ 700 kg 二号培养基或三号培养基的 1 t 种子罐中，在 33 ～ 38°C 的温度下、保持 60 ～ 150 r/min 的转速和 0.5 ～ 3 m3/h 的通气量，培养 2 ～ 5 h ，得到菌液 E ；
    其中，所述三号培养基包括： 5 ～ 20 g/L 的黄豆粕粉、 5 ～ 15g/L 的麸皮、 0 ～ 5 g/L 的 NaCl 、 0 ～ 0.1 g/L 的 K2HPO4 、 0 ～ 0.1 g/L 的 (NH4)2SO4 。
  5. 根据权利要求 4 所述的 苎麻生物脱胶方法，其特征在于，所述 稀释步骤进一步 包括：
    将水注入发酵罐中，保温 34 ～ 37°C ，调节 pH 值为 6.8 ～ 7.5 ，加入所述菌液 E 并混和均匀得到脱胶菌液 F ，所述菌液 E 占脱胶菌液 F 的体积分数为 2% ～ 6% 。
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Publication number Priority date Publication date Assignee Title
CN108823192A (zh) * 2018-06-08 2018-11-16 山东宸青生物科技有限公司 麻类作物纤维精炼酶的生产方法
CN109694836A (zh) * 2019-02-25 2019-04-30 中国农业科学院麻类研究所 一种麻类生物脱胶用固态菌剂的制备方法
CN111519260A (zh) * 2020-03-31 2020-08-11 东华大学 一种麻纤维厌氧生物脱胶罐
CN115029795A (zh) * 2022-07-14 2022-09-09 杭州惠邦纺织有限公司 苎麻纤维吸湿快干面料及其制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1177003A (zh) * 1997-03-19 1998-03-25 武汉大学 新的嗜碱芽孢杆菌菌株及其在苎麻脱胶中的应用
CN1371990A (zh) * 2001-09-26 2002-10-02 山东大学 一株产韧皮纤维脱胶酶的菌株及其在苎麻、大麻脱胶上的应用
CN101285211A (zh) * 2008-06-05 2008-10-15 黑龙江省科学院亚麻综合利用研究所 麻类作物微生物温水沤麻方法
CN101654660A (zh) * 2009-07-14 2010-02-24 东华大学 具有苎麻脱胶活性的枯草芽孢杆菌菌株、其制备和应用
CN101838856A (zh) * 2010-05-14 2010-09-22 华中科技大学 在线苎麻生物脱胶方法
KR101186905B1 (ko) * 2012-05-08 2012-10-02 박선규 바실러스 코아굴란스 에이치피-10 균주를 이용한 인피섬유의 정련방법

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1177003A (zh) * 1997-03-19 1998-03-25 武汉大学 新的嗜碱芽孢杆菌菌株及其在苎麻脱胶中的应用
CN1371990A (zh) * 2001-09-26 2002-10-02 山东大学 一株产韧皮纤维脱胶酶的菌株及其在苎麻、大麻脱胶上的应用
CN101285211A (zh) * 2008-06-05 2008-10-15 黑龙江省科学院亚麻综合利用研究所 麻类作物微生物温水沤麻方法
CN101654660A (zh) * 2009-07-14 2010-02-24 东华大学 具有苎麻脱胶活性的枯草芽孢杆菌菌株、其制备和应用
CN101838856A (zh) * 2010-05-14 2010-09-22 华中科技大学 在线苎麻生物脱胶方法
KR101186905B1 (ko) * 2012-05-08 2012-10-02 박선규 바실러스 코아굴란스 에이치피-10 균주를 이용한 인피섬유의 정련방법

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PANG, ZONGWEN ET AL.: "Study on Ramie Bio-degumming Technology", GUANGXI TEXTILE SCIENCE & TECHNOLOGY, vol. 35, no. 01 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108823192A (zh) * 2018-06-08 2018-11-16 山东宸青生物科技有限公司 麻类作物纤维精炼酶的生产方法
CN109694836A (zh) * 2019-02-25 2019-04-30 中国农业科学院麻类研究所 一种麻类生物脱胶用固态菌剂的制备方法
CN111519260A (zh) * 2020-03-31 2020-08-11 东华大学 一种麻纤维厌氧生物脱胶罐
CN115029795A (zh) * 2022-07-14 2022-09-09 杭州惠邦纺织有限公司 苎麻纤维吸湿快干面料及其制备方法
CN115029795B (zh) * 2022-07-14 2024-01-26 杭州惠邦纺织有限公司 苎麻纤维吸湿快干面料及其制备方法

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